目的 探讨蜈蚣提取物（SSE）对人脐静脉内皮细胞（HUVECs）增殖和凋亡的影响及其与磷脂酰肌醇3激酶（PI3K）/蛋白激酶B（Akt）信号通路的关系。方法 MTT法检测SSE（2、4、8、10、20、40、80、100 mg·mL^-1）、H₂O₂（0.2、0.4、0.6、0.8、1.0 mmol·L^-1）对HUVECs活力的影响，筛选后续造模及给药浓度；采用H₂O₂（0.6 mmol·L^-1）建立HUVECs的细胞损伤模型，设置对照和SSE 10、50、100 mg·mL^-1组。MTT法检测细胞活力；流式细胞术分析细胞凋亡率；Western blotting检测p-PI3K/PI3K、p-Akt/Akt及血管内皮生长因子（VEGF）、缺氧诱导因子-1α（HIF-1α）、B细胞淋巴瘤-2（Bcl-2）、Bcl-2相关X蛋白（Bax）表达；使用PI3K特异性抑制剂LY294002干预以验证通路相关性。结果 与对照组相比，SSE在≤100 mg·mL^-1时对HUVECs无显著细胞毒性； 0.4～1.0 mmol·L^-1的H₂O₂显著降低了HUVECs的细胞存活率（P＜0.01），且呈浓度相关性，0.6 mmol·L^-1 H₂O₂处理组的细胞活力约为65%。与模型组相比，SSE 100 mg·mL^-1显著促进HUVECs增殖（P＜0.01），抑制细胞凋亡（P＜0.01），下调Bax表达（P＜0.01），上调Bcl-2、HIF-1α和VEGF水平（P＜0.01），并提高p-Akt/Akt、p-PI3K/PI3K（P＜0.01）。LY294002处理可显著削弱上述效应（P＜0.01）。结论 SSE可通过激活VEGF-PI3K/Akt信号通路促进HUVECs增殖并抑制其凋亡。

Objective To investigate the effects of Scolopendra subspinipes extract (SSE) on proliferation and apoptosis of human umbilical vein endothelial cells (HUVECs) and its association with the phosphatidylinositol 3-kinase (PI3K)/protein kinase B (Akt) signaling pathway. Methods MTT assay was used to evaluate the effects of SSE (2, 4, 8, 10, 20, 40, 80, 100 mg·mL^-1) and H₂O₂ (0.2, 0.4, 0.6, 0.8, 1.0 mmol·L^-1) on HUVEC viability, to screen suitable concentrations for subsequent model establishment and drug administration. An H₂O₂ (0.6 mmol·L^-1)-induced HUVECs injury model was established, followed by treatment with SSE at 10, 50, and 100 mg·mL^-1. Cell viability was assessed by MTT assay; Apoptosis was analyzed by flow cytometry; Protein expression of p-PI3K/PI3K, p-Akt/Akt ratios, vascular endothelial growth factor (VEGF), hypoxia-inducible factor-1α (HIF-1α), B-cell lymphoma-2 (Bcl-2), and Bcl- 2-associated X (Bax) was detected by Western blotting. The PI3K-specific inhibitor LY294002 was used to confirm pathway involvement. Results Compared with control group, SSE showed no significant cytotoxicity to HUVECs at concentrations ≤ 100 mg·mL^-1. 0.4—1.0 mmol·L^-1 of H₂O₂ significantly reduced the cell viability of HUVECs (P<0.01), and the effect was concentration-dependent. The cell viability of the 0.6 mmol·L^-1 H₂O₂ treatment group was approximately 65%. Compared with the model group, SSE at 100 mg·mL^-1 significantly promoted HUVEC proliferation (P<0.01), inhibited apoptosis (P<0.01), downregulated Bax (P<0.01), and upregulated Bcl-2, HIF-1α, and VEGF (P<0.01), along with increased p-Akt/Akt and p-PI3K/PI3K ratio (P<0.01). These effects were markedly attenuated by LY294002 pretreatment. Conclusion SSE promotes proliferation and suppresses apoptosis in HUVECs via activation of the VEGF-PI3K/Akt signaling pathway, and this effect relies on PI3K/Akt pathway activation.

R965

国家自然科学基金项目（82304470）；湖北省宜昌市医疗卫生科研项目（A15301-17）